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Quantitative acetaminophen analytics

a technology of acetaminophen and analytics, applied in the field of quantitative acetaminophen analytics, can solve the problems of affecting measurement reliability, liver and kidney damage or even death, toxic phenols used in the named tests,

Pending Publication Date: 2020-11-19
DIASYS DIAGNOSTIC SYST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention is a method for quantitatively determining the amount of acetaminophen in a sample. The method involves hydrolyzing acetaminophen with a specific compound, and then reacting the resulting p-aminophenol with a compound of general formula (III) to form a compound of general formula (IV). The amount of compound (IV) is then photometrically determined, allowing for the quantitative determination of acetaminophen in the sample. The method can be used with liquid samples, such as body fluids, and can be carried out without pre-treatment or purification of the sample. The method is advantageously carried out using a specific amidohydrolase enzyme. The compound of general formula (III) contains a negatively charged sulfonate group, which makes it very soluble in polar solvents. The method is particularly useful for analyzing samples containing acetaminophen.

Problems solved by technology

When taken as an overdose, acetaminophen can cause severe liver and kidney damage or even death.
The phenols used in the named tests are toxic and their stability is partly limited, which can impair the measurement reliability.
In any case, the requisite measurement accuracy cannot be reproduced over the entire diagnostically relevant range with sufficient reliability in some cases.
Although both gas-liquid chromatography and high-performance liquid chromatography (HPLC) represent reliable and accurate determination methods, they require a correspondingly large amount of effort and cost-intensive chromatography equipment.
Nevertheless, interactions as well as spectral and chemical interferences with other biomolecules and other therapeutic reagents lead not infrequently to incorrect indications.
However, these coupling agents do not address the problem of interactions with N-acetylcysteine (NAC), which can be contained in a sample to be tested in particular when it has been given as antidote in the case of an already diagnosed acetaminophen poisoning.
Furthermore, the systems which operate with the coupling agents known from the state of the art require relatively large sample volumes, which are, however, not always available in particular in the case of paediatric patient samples as here, in particular in the state of acute acetaminophen poisoning, only very small quantities of blood can be taken.

Method used

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  • Quantitative acetaminophen analytics
  • Quantitative acetaminophen analytics
  • Quantitative acetaminophen analytics

Examples

Experimental program
Comparison scheme
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examples

1. Preliminary Tests

[0068]In order to be able to achieve a still greater precision, a still greater sensitivity and still fewer interactions and fewer spectral and chemical interferences with other compounds contained in the sample for the method according to the invention for quantitatively determining the amount of acetaminophen, different compounds of general formula (III) were tested as coupling agent for the reaction with the p-aminophenol formed from the hydrolysis of acetaminophen.

[0069]For this purpose, a sample was used the concentration of which was 100 mg / L acetaminophen, and, after the hydrolytic cleavage by aryl acylamidase, the reaction with compounds Nos. 1 to 7 specified in the following table was effected.

Abs.No.NameTypeR1R2R3R4(λ = 700 nm)1ALPSquinolineHHC2H5C3H5SO3Na0.082ADPSquinolineOCH3HC2H5C3H6SO3Na0.593TOOSquinolineCH3HC2H5C3H5(OH)SO3Na0.134DAOSquinolineOCH3OCH3C2H5C3H5(OH)SO3Na0.275referenceo-cresol————0.01

[0070]Compound No. 1 does not have the constitution r...

embodiment example

2. Embodiment Example

[0075]In the embodiment described here, the different reagents used for the reaction are provided in the form of a reagent kit with the following solutions, which are stored in separate containers. Here R1 denotes a first reagent solution and R2 denotes a second reagent solution.

[0076]R1 contains, in aqueous solution,[0077]periodate (1.88 mmol / L)[0078]N-ethylmaleimide (10 mM)

[0079]and has a phosphate-buffered pH of 10.65.

[0080]R2 contains, in aqueous solution,[0081]aryl acylamidase (7 u / ml)[0082]ADPS (10 mmol / L)

[0083]and has a phosphate-buffered pH of 7.00.

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Abstract

A method for quantitatively determining the amount of acetaminophen in a sample with greater precision, greater sensitivity and fewer interactions and fewer spectral and chemical interferences with other compounds contained in the sample. The method includes acetaminophen being hydrolyzed and the resulting p-minophenol being reacted with a compound of general formula (III)in the presence of an oxidant, wherein R1 and R2, independently of one another, are selected from H, —CH3, and —OCH3, R3 is —C2H5 and R4 is a C1-4 alkyl moiety with a terminal sulfonate group, with the proviso that at least one of R1 and R2 is —OCH3 and / or R4 additionally has at least one OH substituent, and then the amount of the compound of general formula (IV) in the reaction mixture being photometrically determined.

Description

SUBJECT OF THE INVENTION[0001]The invention relates to a method for quantitatively determining the amount of acetaminophen in a sample and a reagent set for use in the method according to the invention.BACKGROUND OF THE INVENTION[0002]Acetaminophen is the common name in the USA for the active ingredient N-(4-hydroxyphenyl)acetamide), known above all in Europe by the name paracetamol, with formula (I)[0003]Acetaminophen is a widely-used active ingredient, which is used in many non-prescription and prescription-only preparations due to its analgesic and antipyretic properties. When taken as an overdose, acetaminophen can cause severe liver and kidney damage or even death.[0004]After an acute acetaminophen overdose, the patient often displays only few or no symptoms at all at an early stage. The only reliable early indicator for a diagnosis is the quantitative measurement of the acetaminophen concentration in the blood, blood serum or blood plasma. Clinical evidence of liver and kidney...

Claims

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Application Information

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Patent Type & Authority Applications(United States)
IPC IPC(8): C12Q1/34G01N33/94G01N33/52
CPCG01N33/9486C12Q1/34G01N2333/98G01N33/52C07C213/02C07C249/02C07C215/76C07C251/22
Inventor GRIMMLER, MATTHIASTHÖNGES, DETLEFSCHU, PIAMENZENBACH, CAROLINKRECKEL, LAURA
Owner DIASYS DIAGNOSTIC SYST